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Plasmonic materials can generate strong electromagnetic fields to boost the Raman scattering of surrounding molecules, known as surface-enhanced Raman scattering. However, these electromagnetic fields are heterogeneous, with only molecules located at the 'hotspots', which account for ≈ 1% of the surface area, experiencing efficient enhancement. Herein, we propose patterned plasmonic trimers, consisting of a pair of plasmonic dimers at the bilateral sides and a trap particle positioned in between, to address this challenge. The trimer configuration selectively directs probe molecules to the central traps where 'hotspots' are located through chemical affinity, ensuring a precise spatial overlap between the probes and the location of maximum field enhancement. We investigate the Raman enhancement of the Au@Al2O3-Au-Au@Al2O3 trimers, achieving a detection limit of 10-14 M of 4-methylbenzenethiol, 4-mercaptopyridine, and 4-aminothiophenol. Moreover, single-molecule SERS sensitivity is demonstrated by a bi-analyte method. Benefiting from this sensitivity, our approach is employed for the early detection of lung tumors using fresh tissues. Our findings suggest that this approach is sensitive to adenocarcinoma but not to squamous carcinoma or benign cases, offering insights into the differentiation between lung tumor subtypes.
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Oro , Neoplasias Pulmonares , Nanopartículas del Metal , Espectrometría Raman , Espectrometría Raman/métodos , Neoplasias Pulmonares/diagnóstico , Oro/química , Humanos , Nanopartículas del Metal/química , Compuestos de Sulfhidrilo/química , Compuestos de Anilina/química , Adenocarcinoma/diagnóstico , Límite de Detección , Piridinas/químicaRESUMEN
Environmental concerns are driving interest in postpetroleum synthetic textiles produced from microbial and fungal sources. Bacterial cellulose (BC) is a promising sustainable leather alternative, on account of its material properties, low infrastructure needs and biodegradability. However, for alternative textiles like BC to be fully sustainable, alternative ways to dye textiles need to be developed alongside alternative production methods. To address this, we genetically engineer Komagataeibacter rhaeticus to create a bacterial strain that grows self-pigmenting BC. Melanin biosynthesis in the bacteria from recombinant tyrosinase expression achieves dark black coloration robust to material use. Melanated BC production can be scaled up for the construction of prototype fashion products, and we illustrate the potential of combining engineered self-pigmentation with tools from synthetic biology, through the optogenetic patterning of gene expression in cellulose-producing bacteria. With this study, we demonstrate that combining genetic engineering with current and future methods of textile biofabrication has the potential to create a new class of textiles.
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Broad-spectrum antiviral platforms are always desired but still lack the ability to cope with the threats to global public health. Herein, we develop a poly aptamer encoded DNA nanocatcher platform that can trap entire virus particles to inhibit infection with a broad antiviral spectrum. Ultralong single-stranded DNA (ssDNA) containing repeated aptamers was synthesized as the scaffold of a nanocatcher via a biocatalytic process, wherein mineralization of magnesium pyrophosphate on the ssDNA could occur and consequently lead to the formation of nanocatcher with interfacial nanocaves decorated with virus-binding aptamers. Once the viruses were recognized by the apatmers, they would be captured and trapped in the nanocaves via multisite synergistic interactions. Meanwhile, the size of nanocatchers was optimized to prevent their cellular uptake, which further guaranteed inhibition of virus infection. By taking SARS-CoV-2 variants as a model target, we demonstrated the broad virus-trapping capability of a DNA nanocatcher in engulfing the variants and blocking the infection to host cells.
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Aptámeros de Nucleótidos , Virus , Aptámeros de Nucleótidos/farmacología , ADN de Cadena Simple , Antivirales/farmacologíaRESUMEN
The biomolecule deoxyribonucleic acid (DNA), which acts as the carrier of genetic information, is also regarded as a block copolymer for the construction of biomaterials. DNA hydrogels, composed of three-dimensional networks of DNA chains, have received considerable attention as a promising biomaterial due to their good biocompatibility and biodegradability. DNA hydrogels with specific functions can be prepared via assembly of various functional sequences containing DNA modules. In recent years, DNA hydrogels have been widely used for drug delivery, particularly in cancer therapy. Benefiting from the sequence programmability and molecular recognition ability of DNA molecules, DNA hydrogels prepared using functional DNA modules can achieve efficient loading of anti-cancer drugs and integration of specific DNA sequences with cancer therapeutic effects, thus achieving targeted drug delivery and controlled drug release, which are conducive to cancer therapy. In this review, we summarized the assembly strategies for the preparation of DNA hydrogels on the basis of branched DNA modules, hybrid chain reaction (HCR)-synthesized DNA networks and rolling circle amplification (RCA)-produced DNA chains, respectively. The application of DNA hydrogels as drug delivery carriers in cancer therapy has been discussed. Finally, the future development directions of DNA hydrogels in cancer therapy are prospected.
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With the growing development of the Internet of things, wearable electronic devices have been extensively applied in civilian and military fields. As an essential component of data transmission in wearable electronics, a flexible antenna is one of the key aspects of research. Conventional metal antennas suffer from a large skin depth, and cannot satisfy the requirements of wearable electronics such as light weight, flexibility, and thinness. Recently, a group of two-dimensional metallic metal carbides (named MXenes) have been explored as building blocks for high-performance flexible antennas with excellent flexibility and superior mechanical strength. The appearance of hydrophilic functional groups at the surface of a MXene allows simple, scalable, and environmentally friendly manufacturing of MXene-based antennas. In this minireview, some pioneering works of MXene-based flexible radio frequency components are summarized, and the existing bottlenecks and the future trends of this promising field are discussed.
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As an emerging class of two-dimensional (2D) materials, MBenes show enormous potential for optoelectronic applications. However, their use in molecular sensing as surface-enhanced Raman scattering (SERS)-active material is unknown. Herein, for the first time, we develop a brand-new high-performance MBene SERS platform. Ordered vacancy-triggered highly sensitive SERS platform with outstanding signal uniformity based on a 2D Mo4/3B2 MBene material was designed. The 2D Mo4/3B2 MBene presented superior SERS activity to most of the semiconductor SERS substrates, showing a remarkable Raman enhancement factor of 3.88 × 106 and an ultralow detection limit of 1 × 10-9 M. The underlying SERS mechanism is revealed from systematic experiments and density functional theory calculations that the ultrahigh SERS sensitivity of 2D Mo4/3B2 MBene is derived from the efficient photoinduced charge transfer process between MBene substrates and adsorbed molecules. The abundant electronic density of states near the Fermi level of 2D Mo4/3B2 MBene enables its Raman enhancement by a factor of 100 000 times higher than that of the bulk MoB. Consequently, the 2D Mo4/3B2 MBene could accurately detect various trace chemical analytes. Moreover, with ordered metal vacancies in the 2D Mo4/3B2 MBene, uniform charge transfer sites are formed, resulting in an outstanding signal uniformity with a relative standard deviation down to 6.0%. This work opens up a new horizon for the high-performance SERS platform based on MBene materials, which holds great promise in the field of chemical sensing.
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Bacteria proficient at producing cellulose are an attractive synthetic biology host for the emerging field of Engineered Living Materials (ELMs). Species from the Komagataeibacter genus produce high yields of pure cellulose materials in a short time with minimal resources, and pioneering work has shown that genetic engineering in these strains is possible and can be used to modify the material and its production. To accelerate synthetic biology progress in these bacteria, we introduce here the Komagataeibacter tool kit (KTK), a standardized modular cloning system based on Golden Gate DNA assembly that allows DNA parts to be combined to build complex multigene constructs expressed in bacteria from plasmids. Working in Komagataeibacter rhaeticus, we describe basic parts for this system, including promoters, fusion tags, and reporter proteins, before showcasing how the assembly system enables more complex designs. Specifically, we use KTK cloning to reformat the Escherichia coli curli amyloid fiber system for functional expression in K. rhaeticus, and go on to modify it as a system for programming protein secretion from the cellulose producing bacteria. With this toolkit, we aim to accelerate modular synthetic biology in these bacteria, and enable more rapid progress in the emerging ELMs community.
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Celulosa , Ingeniería Genética , Celulosa/genética , Clonación Molecular , Plásmidos/genética , Biología SintéticaRESUMEN
As a type of micro flat loop heat pipe, s-UTLHP (silicon-based ultra-thin loop heat pipe) is of great significance in the field of micro-scale heat dissipation. To prove the feasibility of s-UTLHP with high heat flux in a narrow space, it is necessary to study its heat transfer mechanism visually. In this paper, a structural design of s-UTLHP was proposed, and then, to realize the working fluid charging and visual experiment, an experimental system including a holding module, heating module, cooling module, data acquisition module, and vacuum chamber was proposed. Deionized water was selected as a working fluid in the experiment. The overall and micro phenomena of s-UTLHP during startup, as well as the evaporation and condensation phenomena of s-UTLHP during stable operation, were observed and analyzed. Finally, the failure phenomenon of s-UTLHP was analyzed, and several solutions were proposed. The observed phenomena and experimental conclusions can provide references for further related experimental research.
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The ever-increasing demand for carbon fibre reinforced polymers (CFRP) and stringent environmental legislation have driven the research into recycling and reusing the CFRP waste. This paper presents a mechanical recycling process of CFRP and the application of the recyclates as reinforcement for epoxy foams. The CFRP was mechanically processed using a jet mill. Up to 10 wt% of the CFRP recyclates, without separation of fibre-rich portion and resin-rich portion, was added into epoxy foams. The compressive modulus and strength of the epoxy foams increased from 288 MPa and 7.0 MPa, respectively, to 1060 MPa and 22.8 MPa, respectively, accompanied with an increase in foam density from 0.37 g cm-3 to 0.68 g cm-3. Consequently, the specific compressive modulus and strength (normalised against density) increased from 789 MPa cm3 g-1 and 19.1 MPa cm3 g-1 for unreinforced foam to 1563 MPa cm3 g-1 and 33.5 MPa cm3 g-1 for CFRP recyclates reinforced foam, representing a 98% and 75% improvement, respectively. These results demonstrate that the CFRP recyclates have excellent reinforcing ability for epoxy foams.
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Fibra de Carbono , Polímeros , ReciclajeRESUMEN
The biodegradation of polyacrylamide (PAM) includes the hydrolysis of amino groups and cleavage of the carbon chain; however, the effect of molecular weight on the biodegradation needs further investigations. In this study, biodegradation of low molecular weight PAM (1.6 × 106 Da) was evaluated in two aerobic (25 °C and 40 °C) and two anaerobic (35 °C and 55 °C) reactors over 100 days. The removal of the low molecular weight PAM (52.0-52.6%) through the hydrolysis of amino groups by anaerobic treatment (35 °C and 55 °C) was much higher than that of the high molecular weight (2.2 × 107 Da, 11.2-17.0%) observed under the same conditions. The molecular weight was reduced from 1.6 × 106 to 6.45-7.42 × 105 Da for the low molecular weight PAM, while the high molecular weight PAM declined from 2.2 × 107 to 3.76-5.87 × 106 Da. The results showed that the amino hydrolysis of low molecular weight PAM is easier than that of the high molecular weight one, while the cleavage of its carbon chain is still difficult. The molecular weights of PAM in the effluents from the two aerobic reactors (25 °C and 40 °C) were further reduced to 4.31 × 105 and 5.68 × 105 Da by the biofilm treatment, respectively. The results would be useful for the management of wastewater containing PAM.
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Resinas Acrílicas , Reactores Biológicos , Anaerobiosis , Biodegradación Ambiental , Peso MolecularRESUMEN
Due to the severe restrictions imposed by legislative frameworks, the removal of polyacrylamide (PAM) rapidly and effectively from produced wastewater in offshore oilfields before discharge is becoming an urgent challenge. In this study, a novel advanced oxidation process based on plasma operated in the gas-liquid interface was used to rapidly decompose PAM, and multiple methods including viscometry, flow field-flow fractionation multi-angle light scattering, UV-visible spectroscopy, and attenuated total reflectance-Fourier transform infrared spectroscopy were used to characterize the changes of PAM. Under a discharge voltage of 25â¯kV and pHâ¯7.0, the PAM concentration decreased from 100 to 0â¯mg/L within 20â¯min and the total organic carbon (TOC) decreased from 49.57 to 1.23â¯mg/L within 240â¯min, following zero-order reaction kinetics. Even in the presence of background TOC as high as 152.2â¯mg/L, complete removal of PAM (100â¯mg/L) was also achieved within 30â¯min. The biodegradability of PAM improved following plasma treatment for 120â¯min. Active species (such as O3 and H2O2) were produced in the plasma. Hydroxyl radical was demonstrated to play an important role in the degradation of PAM due to the inhibitory effect observed after the addition of an ·OH scavenger, Na2CO3. Meanwhile, the release of ammonia and nitrate nitrogen confirmed the cleavage of the acylamino group. The results of this study demonstrated that plasma, with its high efficiency and chemical-free features, is a promising technology for the rapid removal of PAM.
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Resinas Acrílicas/química , Eliminación de Residuos Líquidos/métodos , Aguas Residuales/química , Contaminantes Químicos del Agua/química , Resinas Acrílicas/análisis , Biodegradación Ambiental , Peróxido de Hidrógeno/química , Radical Hidroxilo , Cinética , Yacimiento de Petróleo y Gas , Oxidación-Reducción , Contaminantes Químicos del Agua/análisisRESUMEN
Disparate roles exist for tumor-associated macrophages in breast cancer growth and progression. The aim of this study was to explore the influence of induced macrophages on the growth of breast cancer cells. THP-1 monocytes were differentiated to macrophages using phorbol 12-myristate 13-acetate. The effect of the medium from THP-1 monocytes or macrophage-conditioned medium (MφCM) on MCF-7 (estrogen receptor and progesterone-positive positive) and MDA-MB-231 (MB; triple-negative) breast cancer cells was determined at 24â¯h, 48â¯h and 72â¯h. Assays were conducted for cell viability, apoptosis, proliferation and cell phenotype, and quantitative real-time polymerase chain reaction (qRT-PCR) for expression of associated genes. MφCM inhibited proliferation of MCF-7 and MB cells in a time-dependent manner and, in particular, decreased viability of MCF-7 cells. MφCM induced a markedly vacuolated phenotype in MCF-7 increased apoptosis in MCF-7 cells, but correlative changes in Bcl-2 or Bax were absent. A multifold and significant reduction in anti-apoptotic Bcl-2 in MB cells was not matched by increased apoptosis. The cell cycle inhibitor CDKN1A was increased in both cell lines, but PCNA decreased only in MB cells. Senescence-associated galactosidase beta-1 (GLB1) mRNA was decreased in MCF-7 cells (48 and 72â¯h) but increased in MB cells (72â¯h). Increased expression of interleukin-6 (IL-6) and IL-8 was seen in both cell lines, and increased tumor necrosis factor- α was seen at 24â¯h for MB and 72â¯h for MCF-7 indicating increased inflammatory responses of the cancer cells. The two breast cancer celllines had different responses to MφCM, mainly involving inhibition rather than stimulation of growth of the cells, stimulation of senescence (MB cells) and increased inflammatory cytokine expression. The estrogen and progesterone receptor status of the cell lines may determine their response to MφCM. The function of the inflammatory cytokines in breast cancer growth remains to be identified.
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Neoplasias de la Mama/metabolismo , Proliferación Celular , Senescencia Celular , Medios de Cultivo Condicionados/metabolismo , Citocinas/metabolismo , Mediadores de Inflamación/metabolismo , Activación de Macrófagos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Acetato de Tetradecanoilforbol/farmacología , Apoptosis , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/metabolismo , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Proliferación Celular/genética , Supervivencia Celular , Senescencia Celular/genética , Citocinas/genética , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Células MCF-7 , Macrófagos/metabolismo , Comunicación Paracrina , Fenotipo , Transducción de Señal , Células THP-1 , Factores de Tiempo , Neoplasias de la Mama Triple Negativas/genética , Neoplasias de la Mama Triple Negativas/metabolismo , Neoplasias de la Mama Triple Negativas/patologíaRESUMEN
The tumor microenvironment determines development and progression of many cancers. Epithelial-mesenchymal transition (EMT) is fundamental to tumor progression and metastasis not only by increasing invasiveness but also by increasing resistance to cell death, senescence, and various cancer therapies; determining inflammation and immune surveillance; and conferring stem cell properties. It does this by enabling polarized epithelial cells to transform into cells with a mesenchymal, and therefore motile, phenotype. Tumor-associated macrophages (TAMs) are key cells of the tumor microenvironment that orchestrate the connection between inflammation and cancer. Activation of EMT often requires crosstalk between cancer cells and components of the local tumor microenvironment, including TAMs. In this review, clinical and experimental evidence is presented for control of TAMs in promoting cancer cell invasion and migration and their interaction with the EMT process in the metastatic cascade. The translational significance of these findings is that the signaling pathways that interconnect TAMs and EMT-modified cancer cells may represent promising therapeutic targets for the treatment of tumor metastasis.
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High molecular weight partially hydrolyzed polyacrylamide (PAM) can be bio-hydrolyzed on the amide side group, however, solid evidence regarding the biological cleavage of its main carbon chain backbone is limited. In this study, viscometry, flow field-flow fractionation multi-angle light scattering (FFF-MALS), and attenuated total reflectance-Fourier transform infrared (ATR-FTIR) analysis were used to investigate the biodegradability of PAM with a nominal molecular weight of 2 × 107 Da (Da) in two suspended aerobic (25 and 40 °C) and two upflow anaerobic blanket reactors (35 and 55 °C) operated for 470 d under a hydraulic residence time (HRT) of 2 d. Both anaerobic and aerobic biological treatment reduced the viscosity from 2.02 cp in the influent to 1.45-1.60 cp, and reduced the molecular weight of PAM using FFF-MALS from 2.17 × 107 Da to less than one-third its original size. The removals of both the amide group and carbon chain backbone in the PAM molecule were further supported by the FTIR analysis. In comparison with the other conditions, thermophilic anaerobic treatment exhibited higher efficiency for PAM biodegradation. Batch test excluded the influence of temperature on the molecular weight of PAM over the range 25-55 °C, suggesting that cleavage of the main carbon chain backbone was attributed to biological degradation. Our results suggested that high molecular weight PAM was biodegradable, but mineralization did not occur.
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Resinas Acrílicas/química , Modelos Químicos , Anaerobiosis , Biodegradación Ambiental , Reactores Biológicos , Carbono , Hidrólisis , Peso Molecular , Aguas del Alcantarillado , Temperatura , Eliminación de Residuos Líquidos/métodosRESUMEN
Despite the introduction of many novel therapeutics in clinical practice, metastatic renal cell carcinoma (RCC) remains a treatment-resistant cancer. As red and processed meat are considered risk factors for RCC, and a vegetable-rich diet is thought to reduce this risk, research into plant-based therapeutics may provide valuable complementary or alternative therapeutics for the management of RCC. Herein, we present the antiproliferative and antiangiogenic effects of maslinic acid, which occurs naturally in edible plants, particularly in olive fruits, and also in a variety of medicinal plants. Human RCC cell lines (ACHN, Caki-1, and SN12K1), endothelial cells (human umbilical vein endothelial cell line [HUVEC]), and primary cultures of kidney proximal tubular epithelial cells (PTEC) were treated with maslinic acid. Maslinic acid was relatively less toxic to PTEC when compared with RCC under similar experimental conditions. In RCC cell lines, maslinic acid induced a significant reduction in proliferation, proliferating cell nuclear antigen, and colony formation. In HUVEC, maslinic acid induced a significant reduction in capillary tube formation in vitro and vascular endothelial growth factor. This study provides a rationale for incorporating a maslinic acid-rich diet either to reduce the risk of developing kidney cancer or as an adjunct to existing antiangiogenic therapy to improve efficacy.
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Brachyury is an important transcription factor of the T-box gene family with an evolutionarily-conserved function in mesoderm development in the embryo. Recent research has demonstrated that, in various human carcinomas, overexpression of Brachyury is associated with epithelial-mesenchymal transition (EMT), tumor metastasis, expression of markers for cancer stem cells, and resistance to chemotherapy and radiotherapy. Brachyury is a diagnostic and prognostic biomarker, and its expression in tumor tissues is associated with increasing tumor grade, stage, invasiveness, metastasis and poor prognosis. Targeting of Brachyury-positive tumor cells may modulate the extent of EMT and stop invasiveness. Fibroblast growth factor, transforming growth factor-ß and other EMT signalling factors are involved in the molecular pathways of Brachyury in tumorigenesis and development. Experimentally, Brachyury knockdown resulted in downregulation of EMT and stem cell markers, formation of tumor spheroids, and invasiveness. Treatment with recombinant yeast-Brachyury vector-based vaccine can activate and expand Brachyury-specific CD4+ and CD8+ T-cells in vitro, with an outcome of lysis of human tumor cells expressing the Brachyury protein. Further understanding of the characteristics of Brachyury and its associated signaling pathways might help in developing novel therapeutic strategies against EMT.
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Carcinoma/metabolismo , Transición Epitelial-Mesenquimal/fisiología , Proteínas Fetales/metabolismo , Proteínas de Dominio T Box/metabolismo , Biomarcadores de Tumor/metabolismo , Carcinoma/patología , Humanos , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , PronósticoRESUMEN
Adsorption of Cr(VI) onto different pretreated zeolites modified with cetylpyridinium chloride (CPC) is investigated using batch studies, Fourier-transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) measurements and scanning electron microscopy (SEM). The results indicate that acidification after alkalization is the most effective pretreatment method, and only alkalization would significantly reduce the sorption capability. This behavior is due to the precipitates, such as CaCO3 and MgCO3, generated after alkalization on zeolite surface interfere the formation of the CPC bilayer, which provides active sites for sorbing Cr(VI). The schematic of the adsorption mechanism is presented. The results indicate that developing a better understanding of the influence of different pretreatments is quite helpful and suggest that the content of Ca/Mg/Fe could be a good indication of the effectiveness of preprocessing.
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Cromo/química , Zeolitas/química , Adsorción , Carbonato de Calcio/química , Cetilpiridinio/química , Magnesio/química , Microscopía Electrónica de Rastreo , Espectroscopía Infrarroja por Transformada de Fourier , Propiedades de Superficie , Tensoactivos/química , Contaminantes Químicos del Agua/química , Difracción de Rayos XRESUMEN
Column-type combined reactors were designed to cultivate micro-aerobic pentachlorophenol (PCP) degrading granular sludge under oxygen-limited conditions (0.1-0.2 mgL(-1)) over 39-day experimental period. Micro-aerobic granular had both anaerobic activity (SMA: 2.34 mMCH4/hg VSS) and aerobic activity (SOUR: 2.21 mMO2/hg VSS). Metabolite analysis results revealed that PCP was sequentially dechlorinated to TCP, DCP, and eventually to MCP. Methanogens were not directly involved in the dechlorination of PCP, but might played a vital role in stabilizing the overall structure of the granule sludge. For Eubacteria, the Shannon Index (2.09 in inoculated granular sludge) increased both in micro-aerobic granular sludge (2.61) and PCP-degradation granular sludge (2.55). However, for Archaea, it decreased from 2.53 to 1.85 and 1.84, respectively. Although the Shannon Index demonstrated slight difference between micro-aerobic granular sludge and PCP-degradation granular sludge, the Principal Component Analysis (PCA) indicated obvious variance of the microbial composition, revealing significant effect of micro-aerobic condition and PCP on microbial community. Furthermore, nucleotide sequencing indicated that the main microorganisms for PCP degradation might be related to Actinobacterium and Sphingomonas. These results provided insights into situ bioremediation of environments contaminated by PCP and had practical implications for the strategies of PCP degradation.
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Consorcios Microbianos , Pentaclorofenol/metabolismo , Aguas del Alcantarillado/microbiología , Biocombustibles/análisis , Electroforesis en Gel de Gradiente Desnaturalizante , Ácidos Grasos Volátiles/análisis , Análisis de Secuencia de ADNRESUMEN
AIM: To observe the anti-tumor activity of dendritic cell (DC)vaccine loaded with multi-epitopes of survivin. METHODS: The recombinant plasmid pPIRESneo3.0-survivin (4)/Th which include four survivin HLA-A2-restricted CD8(+); CTL epitopes and a CD4(+);Th epitope, pPIRESneo3.0-survivin (4) which include four survivin CD8(+); CTL epitopes, were transfected into human dendritic cells respectively. There were five groups, which included survivin(4)/Th group, survivin(4)group, empty plasmid group, untransfected group and T lymphocytes group The expression of CD83 and CD86 on the surface of DCs, the expression of CD4 and CD8a on the surface of T lymphocytes, the apoptotic rates of MCF-7 cells after treated by DC vaccine were measured by flow cytometry; IFN-γ levels of all groups were detected by ELISA and the growth inhibition of MCF-7 cells after being treated with DC vaccine was tested by MTT colorimetry. RESULTS: The results of flow cytometry revealed that high levels CD83 and CD86 were expressed on the surface of DCs; high levels CD4 and CD8a were expressed on the surface of T lymphocytes; the IFN-γ levels in survivin(4)/Th group [(66.50±3.34)ng/L]were significantly higher than that in survivin(4)group[(46.10±1.35)ng/L], empty plasmid group[(25.17±0.32)ng/L], untransfected group [(25.47±0.95)ng/L] or T lymphocytes group[(23.73±0.50)ng/L](P<0.05). The inhibition rate of MCF-7 cells in survivin(4)/Th group was significantly higher than that in survivin(4)group, empty plasmid group, untransfected group or T lymphocytes group(P<0.05). The apoptotic rate of MCF-7 cells in survivin(4)/Th group was (10.63±0.29)% after treated by DC vaccine, which was significantly higher than that in in survivin(4)group, empty plasmid group, untransfected group or T lymphocytes group(P<0.05). CONCLUSION: The DCs vaccine loaded with multi- CD8(+); CTL epitopes of survivin has strong anti-tumor effects. CD4(+); Th cells can promote the anti-tumor activity of CD8(+);CTL.
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Vacunas contra el Cáncer/inmunología , Vacunas contra el Cáncer/farmacología , Células Dendríticas/inmunología , Epítopos/inmunología , Proteínas Inhibidoras de la Apoptosis/inmunología , Vacunas contra el Cáncer/metabolismo , Muerte Celular/efectos de los fármacos , Línea Celular Tumoral , Células Dendríticas/citología , Células Dendríticas/metabolismo , Humanos , Proteínas Inhibidoras de la Apoptosis/metabolismo , Interferón gamma/efectos de los fármacos , Interferón gamma/metabolismo , SurvivinRESUMEN
AIM: To construct a eukaryotic expression vector encoding the multi-epitope fusion protein of human survivin, and express it in human dendritic cells. METHODS: Recombinant cDNA sequence encoding four HLA-A2-restricted CD8+ CTL epitopes and a CD4+ Th epitope was synthesized and cloned into pBluescript II SK (+) vector. After confirmed by sequencing, the cDNA fragment was inserted to eukaryotic expression vector pIRESneo3.0 to generate the recombinant plasmid pPIRESneo3.0-survivin (4)/Th. The pPIRESneo3.0-survivin (4)/Th was then transfected into human dendritic cells and the transfectants were selected for stable expression. RESULTS: The eukaryotic expression vector encoding the multi-epitope fusion protein of survivin was constructed, and successfully transfected into human dendritic cells. CONCLUSION: The eukaryotic expression vector encoding the multi-epitope fusion protein of survivin has been constructed successfully, and stably expressed in human dendritic cells, which provides clues for further research on multi-epitope cancer vaccine.